Project Summary ? PROJECT 3: JI, Floyd Reed The overall goal of this project is to investigate the complex interactions between Wolbachia, other microbiota, and their mosquito-host's gene regulation as a part of an integrated genetic pest-management system. Mosquito-vectored diseases, especially new and emerging ones, remain one of the largest worldwide threats to human health including within the state of Hawai?i. A wide range of strategies has been proposed to address mosquito-vectored disease and the manipulation of Wolbachia, a symbiotic bacterium found naturally in the majority of arthropod species, has emerged as a promising new approach to reduce vector competence and thereby reduce the rate of infectious disease transmission. However, a great deal of uncertainty surrounds the mechanisms by which Wolbachia infection results in altered vector competence. Furthermore, non-Wolbachia microbiota are known to interact with host vector competence, host gene expression, and Wolbachia. These observations suggest a complex three-way interaction between the mosquito host, Wolbachia, and non- Wolbachia microbiota. This proposed project seeks to leverage next generation sequencing technology, genomics, and state-of-the-art data analysis to explore the basis of these complex interactions and to establish the groundwork necessary to construct a predictive mechanistic model of vector competence. This work will be conducted in two exceptionally widespread mosquito species, Culex quinquefasciatus and Aedes albopictus, that vector a large number of human pathogens. It will also analyze the time component of the evolution of these interactions in two ways. Vector competence, using the dengue virus model, will be assayed both ?early? and ?late? after infection with a novel strain of Wolbachia. Also, vector competence will be analyzed with a fully competent (Aedes) and naive (Culex) host to model the dynamics: (i) when a disease causing pathogen switches hosts to a new species and (ii) within a well established host. This work will have direct implications for a major challenge to human health and will provide an immediate resource for the development of therapeutics for vector-borne disease.